Pharmaceutical compositions

ABSTRACT

The present invention provides pharmaceutical compositions comprising at least one polypeptide having GLP-1 activity wherein an effective dose of said pharmaceutical composition comprises 15 mg, 30 mg, 50 mg or 100 mg of said polypeptide having GLP-1 activity. Also provided are methods of administering the pharmaceutical compositions of the invention.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a divisional of U.S. application Ser. No.13/133,976, filed 10 Jun. 2011, which is a 371 of InternationalApplication No. PCT/US2009/067469, filed 10 Dec. 2009, which isincorporated herein by reference. This application also claims priorityto and benefit of U.S. Provisional Application No. 61/121,229, filed 10Dec. 2008, U.S. Provisional Application No. 61/150,909, filed 9 Feb.2009, U.S. Provisional Application No. 61/163,995, filed 27 Mar. 2009and U.S. Provisional Application No. 61/238,723, filed 1 Sep. 2009.

FIELD OF THE INVENTION

The present invention relates to pharmaceutical compositions and methodsfor administering long-lasting hypoglycemic agents and treatmentregimens using compounds having GLP-1 activity and/or GLP-1 agonists.

BACKGROUND

Hypoglycemic agents may be used in the treatment of both type I and typeII diabetes to lower glucose concentration in blood. Insulinotropicpeptides have been implicated as possible therapeutic agents for thetreatment of diabetes. Insulinotropic peptides include, but are notlimited to, incretin hormones, for example, gastric inhibitory peptide(GIP) and glucagon like peptide-1 (GLP-1), as well as fragments,variants, and/or conjugates thereof. Insulinotropic peptides alsoinclude, for example, exendin 3 and exendin 4. GLP-1 is a 36 amino acidlong incretin hormone secreted by the L-cells in the intestine inresponse to ingestion of food. GLP-1 has been shown to stimulate insulinsecretion in a physiological and glucose-dependent manner, decreaseglucagon secretion, inhibit gastric emptying, decrease appetite, andstimulate proliferation of β-cells. In non-clinical experiments GLP-1promotes continued beta cell competence by stimulating transcription ofgenes important for glucose dependent insulin secretion and by promotingbeta-cell neogenesis (Meier, et al. Biodrugs. 2003; 17 (2): 93-102).

In a healthy individual, GLP-1 plays an important role regulatingpost-prandial blood glucose levels by stimulating glucose-dependentinsulin secretion by the pancreas resulting in increased glucoseabsorption in the periphery. GLP-1 also suppresses glucagon secretion,leading to reduced hepatic glucose output. In addition, GLP-1 delaysgastric emptying and slows small bowel motility delaying foodabsorption.

In people with Type II Diabetes Mellitus (T2DM), the normalpost-prandial rise in GLP-1 is absent or reduced (Vilsboll T, et al.,Diabetes. 2001. 50; 609-613). Accordingly, one rationale foradministering exogenous GLP-1, an incretin hormone, or an incretinmimetic, is to enhance, replace or supplement endogenous GLP-1 in orderto increase meal-related insulin secretion, reduce glucagon secretion,and/or slow gastrointestinal motility. Native GLP-1 has a very shortserum half-life (<5 minutes). Accordingly, it is not currently feasibleto exogenously administer native GLP-1 as a therapeutic treatment fordiabetes. Commercially available incretin mimetics such as Exenatide(Byetta®) improve glycemic control by reducing fasting and postprandialglucose concentrations when administered subcutaneously (5 μg or 10 μgBID) to patients with T2DM.

Thus, there is an unmet need for methods of administering hypoglycemicagents wherein the hypoglycaemic agent is administered at an effectiveweekly or monthly dose.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1. SEQ ID NO.: 1.

FIG. 2: Effects of Albiglutide, Exenatide and Placebo on HbA1c and ADAglycemic targets.

FIG. 3: Effects of Albiglutide, Exenatide and Placebo on HbA1c and ADAglycemic targets.

FIG. 4: Time Course of Nausea and Vomiting as Adverse Events amongPatients Receiving Albiglutide, Exenatide and Placebo.

SUMMARY OF THE INVENTION

The present invention provides pharmaceutical compositions comprising atleast one polypeptide having GLP-1 activity wherein an effective dose ofsaid pharmaceutical composition comprises 15 mg, 30 mg, 50 mg or 100 mgof said polypeptide having GLP-1 activity.

The present invention further provides methods of administering at leastone polypeptide having GLP-1 activity to a human comprisingadministering a pharmaceutical composition of the present invention to ahuman.

DEFINITIONS

“GLP-1 agonist composition” as used herein means any composition capableof stimulating the secretion of insulin, or otherwise raising the levelof insulin, including, but not limited to an incretin hormone and anincretin mimetic.

“Incretin hormone” as used herein means any hormone that potentiatesinsulin secretion or otherwise raises the level of insulin in a mammal.One example of an incretin hormone is GLP-1. GLP-1 is an incretinsecreted by intestinal L cells in response to ingestion of food. In ahealthy individual, GLP-1 plays an important role regulatingpost-prandial blood glucose levels by stimulating glucose-dependentinsulin secretion by the pancreas resulting in increased glucoseabsorption in the periphery. GLP-1 also suppresses glucagon secretion,leading to reduced hepatic glucose output. In addition, GLP-1 delaysgastric emptying time and slows small bowel motility delaying foodabsorption. GLP-1 promotes continued beta cell competence by stimulatingtranscription of genes involved in glucose dependent insulin secretionand by promoting beta-cell neogenesis (Meier, et al. Biodrugs 2003; 17(2): 93-102).

“GLP-1 activity” as used herein means one or more of the activities ofnaturally occurring human GLP-1, including but not limited to, reducingblood and/or plasma glucose, stimulating glucose-dependent insulinsecretion or otherwise raising the level or insulin, suppressingglucagon secretion, reducing fructosamine, increases glucose deliveryand metabolism to the brain, delaying gastric emptying, and promotingbeta cell competence, and/or neogenesis. Any of these activities andother activity associated with GLP-1 activity may be caused directly orindirectly by a composition having GLP-1 activity or a GLP-1 agonist. Byway of example, a composition having GLP-1 activity may directly orindirectly stimulate glucose-dependent while the stimulation of insulinproduction may indirectly reduce plasma glucose levels in a mammal.

An “incretin mimetic” as used herein is a compound capable ofpotentiating insulin secretion or otherwise raise the level or insulin.An incretin mimetic may be capable of stimulating insulin secretion,increasing beta cell neogenesis, inhibiting beta cell apoptosis,inhibiting glucagon secretion, delaying gastric emptying and inducingsatiety in a mammal. An incretin mimetic may include, but is not limitedto, any polypeptide which has GLP-1 activity, including but not limitedto, exendin 3 and exendin 4, including any fragments and/or variantsand/or conjugates thereof.

“Hypoglycemic agent” as used herein means any compound or compositioncomprising a compound capable of reducing blood glucose. A hypoglycemicagent may include, but is not limited to, any GLP-1 agonist includingincretin hormones or incretin mimetics, GLP-1 and/or fragment, variantand/or conjugate thereof. Other hypoglycemic agents include, but are notlimited to, drugs that increase insulin secretion (e.g., sulfonylureas(SU) and meglitinides), inhibit GLP-1 break down (e.g., DPP-IVinhibitors), increase glucose utilization (e.g., glitazones,thiazolidinediones (TZDs) and/or pPAR agonists), reduce hepatic glucoseproduction (e.g., metformin), and delay glucose absorption (e.g.,α-glucosidase inhibitors). Examples of sulfonylureas include but are notlimited to acetohexamide, chlorpropamide, tolazamide, glipizide,gliclazide, glibenclamide (glyburide), gliquidone, and glimepiride.Examples of glitazones include, but are not limited to, rosiglitazoneand pioglitazone.

“Polynucleotide(s)” generally refers to any polyribonucleotide orpolydeoxyribonucleotide that may be unmodified RNA or DNA or modifiedRNA or DNA. “Polynucleotide(s)” include, without limitation, single- anddouble-stranded DNA, DNA that is a mixture of single- anddouble-stranded regions or single-, double- and triple-stranded regions,single- and double-stranded RNA, and RNA that is mixture of single- anddouble-stranded regions, hybrid molecules comprising DNA and RNA thatmay be single-stranded or, more typically, double-stranded, ortriple-stranded regions, or a mixture of single- and double-strandedregions. In addition, “polynucleotide” as used herein refers totriple-stranded regions comprising RNA or DNA or both RNA and DNA. Thestrands in such regions may be from the same molecule or from differentmolecules. The regions may include all of one or more of the molecules,but more typically involve only a region of some of the molecules. Oneof the molecules of a triple-helical region often is an oligonucleotide.As used herein, the term “polynucleotide(s)” also includes DNAs or RNAsas described above that comprise one or more modified bases. Thus, DNAsor RNAs with backbones modified for stability or for other reasons are“polynucleotide(s)” as that term is intended herein. Moreover, DNAs orRNAs comprising unusual bases, such as inosine, or modified bases, suchas tritylated bases, to name just two examples, are polynucleotides asthe term is used herein. It will be appreciated that a great variety ofmodifications have been made to DNA and RNA that serve many usefulpurposes known to those of skill in the art. The term“polynucleotide(s)” as it is employed herein embraces such chemically,enzymatically or metabolically modified forms of polynucleotides, aswell as the chemical forms of DNA and RNA characteristic of viruses andcells, including, for example, simple and complex cells.“Polynucleotide(s)” also embraces short polynucleotides often referredto as oligonucleotide(s).

“Polypeptide” refers to any peptide or protein comprising two or moreamino acids joined to each other by peptide bonds or modified peptidebonds, i.e., peptide isosteres. “Polypeptide” refers to both shortchains, commonly referred to as peptides, oligopeptides or oligomers,and to longer chains, generally referred to as proteins. Polypeptidesmay contain amino acids other than the 20 gene-encoded amino acids.“Polypeptides” include amino acid sequences modified either by naturalprocesses, such as posttranslational processing, or by chemicalmodification techniques that are well known in the art. Suchmodifications are well described in basic texts and in more detailedmonographs, as well as in a voluminous research literature.Modifications can occur anywhere in a polypeptide, including the peptidebackbone, the amino acid side-chains and the amino or carboxyl termini.It will be appreciated that the same type of modification may be presentin the same or varying degrees at several sites in a given polypeptide.Also, a given polypeptide may contain many types of modifications.Polypeptides may be branched as a result of ubiquitination, and they maybe cyclic, with or without branching. Cyclic, branched and branchedcyclic polypeptides may result from posttranslation natural processes ormay be made by synthetic methods. Modifications include acetylation,acylation, ADP-ribosylation, amidation, covalent attachment of flavin,covalent attachment of a heme moiety, covalent attachment of anucleotide or nucleotide derivative, covalent attachment of a lipid orlipid derivative, covalent attachment of phosphotidylinositol,cross-linking, cyclization, disulfide bond formation, demethylation,formation of covalent cross-links, formation of cysteine, formation ofpyroglutamate, formylation, gamma-carboxylation, glycosylation, GPIanchor formation, hydroxylation, iodination, methylation,myristoylation, oxidation, proteolytic processing, phosphorylation,prenylation, racemization, selenoylation, sulfation, transfer-RNAmediated addition of amino acids to proteins such as arginylation, andubiquitination. See, for instance, PROTEINS—STRUCTURE AND MOLECULARPROPERTIES, 2nd Ed., T. E. Creighton, W. H. Freeman and Company, NewYork, 1993 and Wold, F., Posttranslational Protein Modifications:Perspectives and Prospects, pgs. 1-12 in POSTTRANSLATIONAL COVALENTMODIFICATION OF PROTEINS, B. C. Johnson, Ed., Academic Press, New York,1983; Seifter, et al., “Analysis for protein modifications andnonprotein cofactors”, Meth. Enzymol. (1990) 182:626-646 and Rattan, etal., “Protein Synthesis: Posttranslational Modifications and Aging”, AnnNY Acad Sci (1992) 663:48-62.

“Variant” as the term is used herein, is a polynucleotide or polypeptidethat differs from a reference polynucleotide or polypeptiderespectively, but retains essential properties. A typical variant of apolynucleotide differs in nucleotide sequence from another, referencepolynucleotide. Changes in the nucleotide sequence of the variant may ormay not alter the amino acid sequence of a polypeptide encoded by thereference polynucleotide. Nucleotide changes may result in amino acidsubstitutions, additions, deletions, fusions and truncations in thepolypeptide encoded by the reference sequence, as discussed below. Atypical variant of a polypeptide differs in amino acid sequence fromanother, reference polypeptide. Generally, differences are limited sothat the sequences of the reference polypeptide and the variant areclosely similar overall and, in many regions, identical. A variant andreference polypeptide may differ in amino acid sequence by one or moresubstitutions, additions, deletions in any combination. A substituted orinserted amino acid residue may or may not be one encoded by the geneticcode. A variant of a polynucleotide or polypeptide may be a naturallyoccurring such as an allelic variant, or it may be a variant that is notknown to occur naturally. Non-naturally occurring variants ofpolynucleotides and polypeptides may be made by mutagenesis techniquesor by direct synthesis. Variants may also include, but are not limitedto, polypeptides or fragments thereof having chemical modification ofone or more of its amino acid side groups. A chemical modificationincludes, but is not limited to, adding chemical moieties, creating newbonds, and removing chemical moieties. Modifications at amino acid sidegroups include, without limitation, acylation of lysine-ε-amino groups,N-alkylation of arginine, histidine, or lysine, alkylation of glutamicor aspartic carboxylic acid groups, and deamidation of glutamine orasparagine. Modifications of the terminal amino group include, withoutlimitation, the des-amino, N-lower alkyl, N-di-lower alkyl, and N-acylmodifications. Modifications of the terminal carboxy group include,without limitation, the amide, lower alkyl amide, dialkyl amide, andlower alkyl ester modifications. Furthermore, one or more side groups,or terminal groups, may be protected by protective groups known to theordinarily-skilled protein chemist.

As used herein “fragment,” when used in reference to a polypeptide, is apolypeptide having an amino acid sequence that is the same as part butnot all of the amino acid sequence of the entire naturally occurringpolypeptide. Fragments may be “free-standing” or comprised within alarger polypeptide of which they form a part or region as a singlecontinuous region in a single larger polypeptide. By way of example, afragment of naturally occurring GLP-1 would include amino acids 7 to 36of naturally occurring amino acids 1 to 36. Furthermore, fragments of apolypeptide may also be variants of the naturally occurring partialsequence. For instance, a fragment of GLP-1 comprising amino acids 7-30of naturally occurring GLP-1 may also be a variant having amino acidsubstitutions within its partial sequence.

As used herein “conjugate” or “conjugated” refers to two molecules thatare bound to each other. For example, a first polypeptide may becovalently or non-covalently bound to a second polypeptide. The firstpolypeptide may be covalently bound by a chemical linker or may begenetically fused to the second polypeptide, wherein the first andsecond polypeptide share a common polypeptide backbone.

As used herein “tandemly oriented” refers to two or more polypeptidesthat are adjacent to one another as part of the same molecule. They maybe linked either covalently or non-covalently. Two or more tandemlyoriented polypeptides may form part of the same polypeptide backbone.Tandemly oriented polypeptides may have direct or inverted orientationand/or may be separated by other amino acid sequences.

As used herein, “reduce” or “reducing” blood or plasma glucose refers toa decrease in the amount of blood glucose observed in the blood of apatient after administration a hypoglycemic agent. Reductions in bloodor plasma glucose can be measured and assessed per individual or as amean change for a group of subjects. Additionally, mean reductions inblood or plasma glucose can be measured and assessed for a group oftreated subjects as a mean change from baseline and/or as a mean changecompared with the mean change in blood or plasma glucose among subjectsadministered placebo.

As used herein “enhancing GLP-1 activity” refers to an increase in anyand all of the activities associated with naturally occurring GLP-1. Byway of example, enhancing GLP-1 activity can be measured afteradministration of at least one polypeptide having GLP-1 activity to asubject and compared with GLP-1 activity in the same subject prior tothe administration of the polypeptide having GLP-1 activity or incomparison to a second subject who is administered placebo.

As used herein “diseases associated with elevated blood glucose”include, but are not limited to, type I and type II diabetes, glucoseintolerance, and hyperglycemia.

As used herein “co-administration” or “co-administering” refers toadministration of two or more compounds or two or more doses of the samecompound to the same patient. Co-administration of such compounds may besimultaneous or at about the same time (e.g., within the same hour) orit may be within several hours or days of one another. For example, afirst compound may be administered once weekly while a second compoundis co-administered daily.

As used herein “maximum plasma concentration” or “Cmax” means thehighest observed concentration of a substance (for example, apolypeptide having GLP-1 activity or a GLP-1 agonist) in mammalianplasma after administration of the substance to the mammal.

As used herein “Area Under the Curve” or “AUC” is the area under thecurve in a plot of the concentration of a substance in plasma againsttime. AUC can be a measure of the integral of the instantaneousconcentrations during a time interval and has the unitsmass×time/volume, which can also be expressed as molarconcentration×time such as nM×day. AUC is typically calculated by thetrapezoidal method (e.g., linear, linear-log). AUC is usually given forthe time interval zero to infinity, and other time intervals areindicated (for example AUC (t1, t2) where t1 and t2 are the starting andfinishing times for the interval). Thus, as used herein “AUC_(0-24h)”refers to an AUC over a 24-hour period, and “AUC_(0-4h)” refers to anAUC over a 4-hour period.

As used herein “weighted mean AUC” is the AUC divided by the timeinterval over which the time AUC is calculated. For instance, weightedmean AUC_(0-24h) would represent the AUC_(0-24h) divided by 24 hours.

As used herein “confidence interval” or “CI” is an interval in which ameasurement or trial falls corresponding to a given probability p wherep refers to a 90% or 95% CI and are calculated around either anarithmetic mean, a geometric mean, or a least squares mean. As usedherein, a geometric mean is the mean of the natural log-transformedvalues back-transformed through exponentiation, and the least squaresmean may or may not be a geometric mean as well but is derived from theanalysis of variance (ANOVA) model using fixed effects.

As used herein the “coefficient of variation (CV)” is a measure ofdispersion and it is defined as the ratio of the standard deviation tothe mean. It is reported as a percentage (%) by multiplying the abovecalculation by 100 (% CV).

As used herein “Tmax” refers to the observed time for reaching themaximum concentration of a substance in plasma of a mammal afteradministration of that substance to the mammal.

As used herein “serum or plasma half life” refers to the time requiredfor half the quantity of a substance administered to a mammal to bemetabolized or eliminated from the serum or plasma of the mammal bynormal biological processes.

As used herein “dose” refers to any amount of therapeutic compound whichmay be administered to a mammal, including a human. An effective dose isa dose of a compound that is in an amount sufficient to induce at leastone of the intended effects of the therapeutic compound. For instance,an effective dose of a GLP-1 agonist would induce at least one type ofGLP-1 activity in a human when administered to a human, such as, but notlimited to increasing insulin production in said human. As is understoodin the art, an effective dose of a therapeutic compound can be measuredby a surrogate endpoint. Thus, by way of another example, an effectivedose of a GLP-1 agonist can be measured by its ability to lower serumglucose in a human.

As used herein “cardiovascular disorder” include, but is not limited to,cardiovascular abnormalities, such as arterio-arterial fistula,arteriovenous fistula, cerebral arteriovenous malformations, congenitalheart defects, pulmonary atresia, and Scimitar Syndrome. Congenitalheart defects include, but are not limited to, aortic coarctation, cortriatriatum, coronary vessel anomalies, crisscross heart, dextrocardia,patent ductus arteriosus, Ebstein's anomaly, Eisenmenger complex,hypoplastic left heart syndrome, levocardia, tetralogy of fallot,transposition of great vessels, double outlet right ventricle, tricuspidatresia, persistent truncus arteriosus, and heart septal defects, suchas aortopulmonary septal defect, endocardial cushion defects,Lutembacher's Syndrome, trilogy of Fallot, ventricular heart septaldefects.

Cardiovascular disorders also include, but are not limited to, chroniccardiac failure, heart disease, such as arrhythmias, carcinoid heartdisease, high cardiac output, low cardiac output, cardiac tamponade,endocarditis (including bacterial), heart aneurysm, cardiac arrest,congestive heart failure, congestive cardiomyopathy, diabeticcardiomyopathy, paroxysmal dyspnea, cardiac edema, heart hypertrophy,congestive cardiomyopathy, left ventricular hypertrophy, rightventricular hypertrophy, post-infarction heart rupture, ventricularseptal rupture, heart valve diseases, myocardial diseases, myocardialischemia, pericardial effusion, pericarditis (including constrictive andtuberculous), pneumopericardium, postpericardiotomy syndrome, pulmonaryheart disease, rheumatic heart disease, ventricular dysfunction,hyperemia, cardiovascular pregnancy complications, Scimitar Syndrome,cardiovascular syphilis, and cardiovascular tuberculosis.

Arrhythmias include, but are not limited to, sinus arrhythmia, atrialfibrillation, atrial flutter, bradycardia, extrasystole, Adams-StokesSyndrome, bundle-branch block, sinoatrial block, long QT syndrome,parasystole, Lown-Ganong-Levine Syndrome, Mahaim-type pre-excitationsyndrome, Wolff-Parkinson-White syndrome, sick sinus syndrome,tachycardias, and ventricular fibrillation. Tachycardias includeparoxysmal tachycardia, supraventricular tachycardia, acceleratedidioventricular rhythm, atrioventricular nodal reentry tachycardia,ectopic atrial tachycardia, ectopic junctional tachycardia, sinoatrialnodal reentry tachycardia, sinus tachycardia, Torsades de Pointes, andventricular tachycardia.

Heart valve diseases include, but are not limited to, aortic valveinsufficiency, aortic valve stenosis, hear murmurs, aortic valveprolapse, mitral valve prolapse, tricuspid valve prolapse, mitral valveinsufficiency, mitral valve stenosis, pulmonary atresia, pulmonary valveinsufficiency, pulmonary valve stenosis, tricuspid atresia, tricuspidvalve insufficiency, and tricuspid valve stenosis.

Myocardial diseases include, but are not limited to, alcoholiccardiomyopathy, diabetic cardiomyopathy, congestive cardiomyopathy,hypertrophic cardiomyopathy, aortic subvalvular stenosis, pulmonarysubvalvular stenosis, restrictive cardiomyopathy, Chagas cardiomyopathy,endocardial fibroelastosis, endomyocardial fibrosis, Kearns Syndrome,myocardial reperfusion injury, and myocarditis.

Myocardial ischemias include, but are not limited to, coronary disease,such as angina pectoris, coronary aneurysm, coronary arteriosclerosis,coronary thrombosis, coronary vasospasm, myocardial infarction andmyocardial stunning.

Cardiovascular diseases also include vascular diseases such asaneurysms, angiodysplasia, angiomatosis, bacillary angiomatosis,Hippel-Lindau Disease, Klippel-Trenaunay-Weber Syndrome, Sturge-WeberSyndrome, angioneurotic edema, aortic diseases, Takayasu's Arteritis,aortitis, Leriche's Syndrome, arterial occlusive diseases, arteritis,enarteritis, polyarteritis nodosa, cerebrovascular disorders, diabeticangiopathies, diabetic retinopathy, embolisms, thrombosis,erythromelaigia, hemorrhoids, hepatic veno-occlusive disease,hypertension, hypotension, ischemia, peripheral vascular diseases,phlebitis, pulmonary veno-occlusive disease, Raynaud's disease, CRESTsyndrome, retinal vein occlusion, Scimitar syndrome, superior vena cavasyndrome, telangiectasia, atacia telangiectasia, hereditary hemorrhagictelangiectasia, varicocele, varicose veins, varicose ulcer, vasculitis,and venous insufficiency.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides pharmaceutical compositions comprising atleast one polypeptide having GLP-1 activity wherein an effective dose ofsaid pharmaceutical composition comprises 15 mg, 30 mg, 50 mg, or 100 mgof said polypeptide having GLP-1 activity. In some aspects, thepolypeptide having GLP-1 activity comprises at least one GLP-1 agonist.GLP-1 agonists can be selected from the group of: incretin hormoneand/or fragment, variant and/or conjugate thereof and incretin mimeticand/or fragment, variant and/or conjugate thereof. Included amongincretin hormones are human GLP-1 and/or fragments, variants and/orconjugates thereof.

An embodiment of the invention comprises a polypeptide that may be, butis not limited to, GLP-1 or a fragment, variant, and/or conjugatethereof. GLP-1 fragments and/or variants and/or conjugates of thepresent invention typically have at least one GLP-1 activity. A GLP-1 ora fragment, variant, and/or conjugate thereof may comprise human serumalbumin. Human serum albumin may be conjugated to the GLP-1 or fragmentand/or variant thereof. Human serum albumin may be conjugated to anincretin hormone (such as GLP-1) and/or incretin mimetic (such asexendin 3 and exendin 4) and/or fragments and/or variants thereofthrough a chemical linker prior to injection or may be chemically linkedto naturally occurring human serum albumin in vivo (see for instance,U.S. Pat. No. 6,593,295 and U.S. Pat. No. 6,329,336, herein incorporatedby reference in their entirety). Alternatively, human serum albumin maybe genetically fused to a GLP-1 and/or fragment and/or variant thereofor other GLP-1 agonist such as exendin-3 or exendin-4 and/or fragmentsand/or variants thereof. Examples of GLP-1 and fragments and/or variantsthereof fused with human serum albumin are provided in the following: WO2003/060071, WO 2003/59934, WO 2005/003296, WO 2005/077042 and U.S. Pat.No. 7,141,547 (herein incorporated by reference in their entirety).

Polypeptides having GLP-1 activity may comprise at least one fragmentand/or variant of human GLP-1. The two naturally occurring fragments ofhuman GLP-1 are represented in SEQ ID NO: 2.

(SEQ ID NO.: 2) 7   8   9   10  11  12  13  14  15  16  17His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-18  19  20  21  22  23  24  25  26  27  28Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-29  30  31  32  33  34  35  36  37 Ile-Ala-Trp-Leu-Val-Lys-Gly-Arg-Xaa wherein: Xaa at position 37 is Gly (hereinafter designated as“GLP-1(7-37)”), or —NH₂ (hereinafter designated as “GLP-1(7-36)”). GLP-1fragments may include, but are not limited to, molecules of GLP-1comprising, or alternatively consisting of, amino acids 7 to 36 of humanGLP-1 (GLP-1(7-36)). Variants of GLP-1 or fragments thereof may include,but are not limited to, one, two, three, four, five or more amino acidsubstitutions in wild type GLP-1 or in the naturally occurring fragmentsof GLP-1 shown in SEQ ID NO.: 2. Variants GLP-1 or fragments of GLP-1may include, but are not limited to, substitutions of an alanine residueanalogous to alanine 8 of wild type GLP-1, such alanine being mutated toa glycine (hereinafter designated as “A8G”) (See for example, themutants disclosed in U.S. Pat. No. 5,545,618, herein incorporated byreference in its entirety).

In some aspects, at least one fragment and variant of GLP-1 comprisesGLP-1(7-36(A8G)) and is genetically fused to human serum albumin. In afurther embodiment, polypeptides of the invention comprise one, two,three, four, five, or more tandemly oriented molecules of GLP-1 and/orfragments and/or variants thereof fused to the N- or C-terminus of humanserum albumin or variant thereof. Other embodiments have such A8Gpolypeptides fused to the N- or C-terminus of albumin or variantthereof. An example of two tandemly oriented GLP-1(7-36)(A8G) fragmentsand/or variants fused to the N-terminus of human serum albumin comprisesSEQ ID NO:1, which is presented in FIG. 1. In another aspect, at leastone fragment and variant of GLP-1 comprises at least twoGLP-1(7-36(A8G)) tandemly and genetically fused to the human serumalbumin. At least two GLP-1(7-36(A8G)) may be genetically fused at theN-terminus of the human serum albumin. At least one polypeptide havingGLP-1 activity may comprise SEQ ID No.: 1.

Variants of GLP-1(7-37) may be denoted for example asGlu²²-GLP-1(7-37)OH which designates a GLP-1 variant in which theglycine normally found at position 22 of GLP-1(7-37)OH has been replacedwith glutamic acid; Val⁸-Glu²²-GLP-1(7-37)OH designates a GLP-1 compoundin which alanine normally found at position 8 and glycine normally foundat position 22 of GLP-1(7-37)OH have been replaced with valine andglutamic acid, respectively. Examples of variants of GLP-1 include, butare not limited to,

Val⁸-GLP-1(7-37)OH Gly⁸-GLP-1(7-37)OH Glu²²-GLP-1(7-37)O—HAsp²²-GLP-1(7-37)OH Arg²²-GLP-1(7-37)OH Lys²²-GLP-1(7-37)OHCys²²-GLP-1(7-37)OH Val⁸-Glu²²-GLP-1(7-37)OH Val⁸-Asp²²-GLP-1(7-37)OHVal⁸-Arg²²-GLP-1(7-37)OH Val⁸-Lys²²-GLP-1(7-37)OHVal⁸-Cys²²-GLP-1(7-37)OH Gly⁸-Glu²²-GLP-1(7-37)OHGly⁸-Asp²²-GLP-1(7-37)OH Gly⁸-Arg²²-GLP-1(7-37)OHGly⁸-Lys²²-GLP-1(7-37)OH Gly⁸-Cys²²-GLP-1(7-37)OH Glu²²-GLP-1(7-36)OHAsp²²-GLP-1(7-36)OH Arg²²-GLP-1(7-36)OH Lys²²-GLP-1(7-36)OHCys²²-GLP-1(7-36)OH Val⁸-Glu²²-GLP-1(7-36)OH Val⁸-Asp²²-GLP-1(7-36)OHVal⁸-Arg²²-GLP-1(7-36)OH Val⁸-Lys²²-GLP-1(7-36)OHVal⁸-Cys²²-GLP-1(7-36)OH Gly⁸-Glu²²-GLP-1(7-36)OHGly⁸-Asp²²-GLP-1(7-36)OH Gly⁸-Arg²²-GLP-1(7-36)OHGly⁸-Lys²²-GLP-1(7-36)OH Gly⁸-Cys²²-GLP-1(7-36)OH Lys²³-GLP-1(7-37)OHVal⁸-Lys²³-GLP-1(7-37)OH Gly⁸-Lys²³-GLP-1(7-37)OH His²⁴-GLP-1(7-37)OHVal⁸-His²⁴-GLP-1(7-37)OH Gly⁸-His²⁴-GLP-1(7-37)OH Lys²⁴-GLP-1(7-37)OHVal⁸-Lys²⁴-GLP-1(7-37)OH Gly⁸-Lys²³-GLP-1(7-37)OH Glu³⁰-GLP-1(7-37)OHVal⁸-Glu³⁰-GLP-1(7-37OH Gly⁸-Glu³⁰-GLP-1(7-37)OH Asp³⁰-GLP-1(7-37)OHVal⁸-Asp³⁰-GLP-1(7-37)OH Gly⁸-Asp³⁰-GLP-1(7-37)OH Gln³⁰-GLP-1(7-37)OHVal⁸-Gln³⁰-GLP-1(7-37)OH Gly⁸-Gln³⁰-GLP-1(7-37)OH Tyr³⁰-GLP-1(7-37)OHVal⁸-Tyr³⁰-GLP-1(7-37)OH Gly⁸-Tyr³⁰-GLP-1(7-37)OH Ser³⁰-GLP-1(7-37)OHVal⁸-Ser³⁰-GLP-1(7-37)OH Gly⁸-Ser³⁰-GLP-1(7-37)OH His³⁰-GLP-1(7-37)OHVal⁸-His³⁰-GLP-1(7-37)OH Gly⁸-His³⁰-GLP-1(7-37)OH Glu³⁴-GLP-1(7-37)OHVal⁸-Glu³⁴-GLP-1(7-37)OH Gly⁸-Glu³⁴-GLP-1(7-37)OH Ala³⁴-GLP-1(7-37)OHVal⁸-Ala³⁴-GLP-1(7-37)OH Gly⁸-Ala³⁴-GLP-1(7-37)OH Gly³⁴-GLP-1(7-37)OHVal⁸-Gly³⁴-GLP-1(7-37)OH Gly⁸-Gly³⁴-GLP-1(7-37)OH Ala³⁵-GLP-1(7-37)OHVal⁸-Ala³⁵-GLP-1(7-37)OH Gly⁸-Ala³⁵-GLP-1(7-37)OH Lys³⁵-GLP-1(7-37)OHVal⁸-Lys³⁵-GLP-1(7-37)OH Gly⁸-Lys³⁵-GLP-1(7-37)OH His³⁵-GLP-1(7-37)OHVal⁸-His³⁵-GLP-1(7-37)OH Gly⁸-His³⁵-GLP-1(7-37)OH Pro³⁵-GLP-1(7-37)OHVal⁸-Pro³⁵-GLP-1(7-37)OH Gly⁸-Pro³⁵-GLP-1(7-37)OH Glu³⁵-GLP-1(7-37)OHGly⁸-Glu³⁵-GLP-1(7-37)OH Val⁸-Ala²⁷-GLP-1(7-37)OHVal⁸-His³⁷-GLP-1(7-37)OH Val⁸-Glu²²-Lys²³-GLP-1(7-37)OHVal⁸-Glu²²-Glu²³-GLP-1(7-37)OH Val⁸-Glu²²-Ala²⁷-GLP-1(7-37)OHVal⁸-Gly³⁴-Lys³⁵-GLP-1(7-37)OH Val⁸-His³⁷-GLP-1-(7-37)OHGly⁸-His³⁷-GLP-1(7-37)OH Val⁸-Glu²²-Ala²⁷-GLP-1(7-37)OHGly⁸-Glu²²-Ala²⁷-GLP-1(7-37)OH Val⁸-Lys²²-Glu²³-GLP-1(7-37)OHGly⁸-Lys²²-Glu²³-GLP-1(7-37)OH• Val⁸-Glu³⁵-GLP-1(7-37)OH

Variants of GLP-1 may also include, but are not limited to, GLP-1 orGLP-1 fragments having chemical modification of one or more of its aminoacid side groups. A chemical modification includes, but is not limitedto, adding chemical moieties, creating new bonds, and removing chemicalmoieties. Modifications at amino acid side groups include, withoutlimitation, acylation of lysine-ε-amino groups, N-alkylation ofarginine, histidine, or lysine, alkylation of glutamic or asparticcarboxylic acid groups, and deamidation of glutamine or asparagine.Modifications of the terminal amino group include, without limitation,the des-amino, N-lower alkyl, N-di-lower alkyl, and N-acylmodifications. Modifications of the terminal carboxy group include,without limitation, the amide, lower alkyl amide, dialkyl amide, andlower alkyl ester modifications. Furthermore, one or more side groups,or terminal groups, may be protected by protective groups known to theordinarily-skilled protein chemist.

GLP-1 fragments or variants may also include polypeptides in which oneor more amino acids have been added to the N-terminus and/or C-terminusof GLP-1(7-37)OH of said fragment or variant. The amino acids in GLP-1in which amino acids have been added to the N-terminus or C-terminus aredenoted by the same number as the corresponding amino acid inGLP-1(7-37)OH. For example, the N-terminus amino acid of a GLP-1compound obtained by adding two amino acids to the N-terminus ofGLP-1(7-37)OH is at position 5; and the C-terminus amino acid of a GLP-1compound obtained by adding one amino acid to the C-terminus ofGLP-1(7-37)OH is at position 38. Thus, position 12 is occupied byphenylalanine and position 22 is occupied by glycine in both of theseGLP-1 compounds, as in GLP-1(7-37)OH. Amino acids 1-6 of a GLP-1 withamino acids added to the N-terminus may be the same as or a conservativesubstitution of the amino acid at the corresponding position ofGLP-1(1-37)OH. Amino acids 38-45 of a GLP-1 with amino acids added tothe C-terminus may be the same as or a conservative substitution of theamino acid at the corresponding position of glucagon or exendin-4.

In another embodiment, the pharmaceutical composition of the presentinvention can administered to a human once daily, once every other day,once every seven days, once every fourteen days, once every four weeks,and/or once every month. In another aspect, the pharmaceuticalcompositions comprises at least 30 mg/mL of SEQ ID NO:1. In anotheraspect, the pharmaceutical composition consists of 30 mg/mL of SEQ IDNO:1, sodium phosphate, trehalose, mannitol, TWEEN 80 and water and ismaintained at pH 7.2. In another aspect, the pharmaceutical compositionconsists of 50 mg/mL of SEQ ID NO:1, sodium phosphate, trehalose,mannitol, TWEEN 80 and water and is maintained at pH 7.2.

In another aspect, methods are provided for administering at least onepolypeptide having GLP-1 activity to a human comprising administering apharmaceutical composition of the invention to a human. Pharmaceuticalcomposition can be administered subcutaneously. Pharmaceuticalcomposition can be administered as a subcutaneous injection selectedfrom the group of: at least one 0.32 mL injection, at least one 0.65 mLinjection, and at least one 1.0 mL injection. In some aspects, thepharmaceutical composition is co-administered in two injections, whichmay be the same dose or may be different doses of the samepharmaceutical composition. The pharmaceutical compositions of thepresent invention may be administered at the same or different injectionsites. Subcutaneous injections of the invention may be administered assingle injections, meaning the entire dose is administered as a singleshot, wherein the entire volume of the shot is administered all at once.A single shot differs from a continuous administration which may beadministered over several minutes and/or hours and/or days. Singleinjections may be administered multiple times, meaning as a single shotonce daily, weekly, every two weeks, monthly and/or more.

In another aspect, the pharmaceutical composition reduces HbA1c in saidhuman and/or serum glucose in said human. The serum half life of said atleast one polypeptide having GLP-1 activity, for example SEQ ID NO:1, isabout 5 days. In another aspect, the human has a disease associated withelevated glucose levels which may include hyperglycemia, diabetes, typeII diabetes. In yet another aspect, the pharmaceutical compositions ofthe present invention cause weight loss in a human when administered toa human.

In another embodiment, pharmaceutical compositions of the presentinvention are administered as monotherapy. In yet another embodiment,the pharmaceutical compositions is co-administered with at least onesecond hypoglycaemic agent. A second hypoglycaemic agent may be selectedfrom: a GLP-1 agonist, incretin hormone, incretin mimetic, agent toincrease insulin secretion, sulfonylurea, meglitinide, acetohexamide,chlorpropamide, tolazamide, glipizide, gliclazide, glibenclamide(glyburide), gliquidone, glimepiride, agent to inhibit GLP-1 break down,DPP-IV inhibitor, agent to increase glucose utilization, glitazones,thiazolidinediones, rosiglitazone, pioglitazone, pPAR agonists, agent toreduce hepatic glucose production, metformin, agent to delay glucoseabsorption, α-glucosidase inhibitor, insulin glargine and/or insulin. Insome aspects, the second hypoglycaemic agent is metformin. In someaspects of the present invention, the polypeptide having GLP-1 activityis administered to said human at an initial dose of 30 mg andsubsequently titrated up to 50 mg. The skilled artisan will understandthat pharmaceutical compositions can be administered to humans who areno longer responding to their current therapy. That is, a subject mayhave a wash-out period from current therapy while concurrently orsequentially starting therapy with a pharmaceutical composition of thepresent invention.

As is understood in the art, various methods may be employed to collect,measure and assess pharmacokinetic data such as active compoundconcentration in blood, plasma and/or other tissue. As is alsounderstood in the art, various methods may be employed to collect,measure and assess various pharmacodynamic data such as, but not limitedto, glucose, insulin, C peptide, glucagon and other biomarker levels inblood and/or plasma and/or other tissue.

A skilled artisan will understand the various methods for measuring andcalculating the pharmacokinetic (for example, but not limited to, Cmax,AUC, Tmax, serum half-life) and pharmacodynamic (for example, but notlimited to, serum and/or plasma blood glucose levels and/or HbA1clevels) parameters described herein. Furthermore, the skilled artisanwill understand the various methods for making statistical comparisons(for example, but not limited to, comparisons of change from baseline topost-treatment and/or comparisons among treatment groups) and/oranalysis of the pharmacokinetic and pharmacodynamic parameters describedherein. Furthermore, the skilled artisan will understand and be able toemploy various other methods for collecting and analyzingpharmacokinetic, pharmacodynamic and other clinical data.

Furthermore, the present invention includes pharmaceutical compositionsas well as methods of making the pharmaceutical compositions and methodsof using the pharmaceutical compositions of the present invention. Thepharmaceutical compositions of the present invention can be used intreatment or prophylactically to treat, prevent and/or prevent theworsening of any symptom of any disease or condition associated withelevated glucose, obesity, cardiovascular disorders, including but notlimited to myocardial infarction, and chronic heart failure, and/ormemory loss.

EXAMPLES

The following examples illustrate various non-limiting aspects of thisinvention. For the following examples, unless noted otherwise, SEQ IDNO.: 1 also referred to herein as Albiglutide (ALB) was formulated as 50mg/mL from a lyophilized form comprising 2.8% mannitol, 4.2% trehalosedihydrate, 0.01% polysorbate 80, 10 to 20 mM phosphate buffer at pH 7.2.Compositions comprising SEQ ID NO.: 1 were diluted with water forinjection as necessary for respective dosing.

Example 1 The Potential of Albiglutide, a Long-Acting GLP-1 Mimetic, inType 2 Diabetes: A Randomized Controlled Trial Exploring Weekly,Biweekly and Monthly Dosing

This study was designed to evaluate the efficacy, safety andtolerability of incremental doses of albiglutide (ALB), a long-actingGLP-1-receptor agonist, administered with 3 timing schedules in type 2diabetic patients inadequately controlled with diet and exercise ormetformin monotherapy. Albiglutide (ALB) consists of a DPP-4-resistantGLP-1 dimer fused to human albumin. With a half-life of ˜5 days, ALB hasthe potential for weekly or less-frequent dosing. In this randomizedmulticenter, double-blind, parallel-group study in 356 type 2 diabeticsubjects with similar mean baseline characteristics (age 53.4 years,diabetes duration 5 years, BMI 32 kg/m², A1C 8.0%), patients receivedsubcutaneous placebo, albiglutide [weekly (4, 15 or 30 mg), every-otherweek (biweekly; 15, 30 or 50 mg) or monthly (50 or 100 mg)], orexenatide as an open-label active reference (twice-daily per labeling inmetformin patients only) over 16 weeks, followed by an 11-week washoutperiod. Main outcome measure was change from baseline A1C, week 16 vsplacebo.

Outcomes studies show that early intervention to improve glycemiccontrol reduces microvascular complications in type 2 diabetes (UKProspective Diabetes Study (UKPDS) (UKPDS 33). Lancet 352:837-853, 1998;Gerstein, et al. N Engl J Med 358:2545-2559, 2008; Patel, et al. N EnglJ Med 358:2560-2572, 2008; and Abrair, et al. Diabetes Obes Metab 2008;DOI: 10.1111/j.1463-1326.2008.00933.x) and may provide long-termmacrovascular benefit (Holma, et al. N Engl J Med 359:1577-1589, 2008).Despite numerous available therapies, over half of patients with type 2diabetes are unable to achieve the American Diabetes Association (ADA)target A1C level (<7%) (Saydah, et al. JAMA 291:335-342, 2004; Saaddine,et al. Ann Intern Med 144(7):465-474, 2006; Ong K L, et al. AnnEpidemiol 18:222-229, 2008). Moreover, weight gain and treatment-inducedhypoglycemic episodes (Carver. Diabetes Educ 32:910-917, 2006 and Kahn,et al. N Engl J Med 355:2427-2443, 2006) are major barriers to achievingglycemic control (Bray G M. Exenatide. Am J Health Syst Pharm63:411-418, 2006). Antidiabetic therapies, based on glucagon-likepeptide-1 (GLP-1), retain the ability of native GLP-1 to stimulateglucose-dependent insulin secretion and suppress inappropriatelyelevated post-meal glucagon secretion (Drucker, et al. Proc Natl AcadSci USA 84:3434-3438, 1987; Kreymann, et al. Lancet 2:1300-1304, 1987).Native GLP-1 also slows gastric emptying and reduces food intake,leading to modest weight loss in patients with T2DM. (Hols, et al.Trends Moled Med 14(4):161-168, 2008). However, native GLP-1 is rapidlyinactivated (half-life: 1-2 minutes) by dipeptidyl peptidase-4 (DPP-4),limiting its therapeutic potential (Deacon, et al. Am J Physiol 271(3 Pt1):E458-E464, 1996). Exenatide (half-life: 2.4 hours) improves glycemiccontrol in combination with metformin, sulfonylurea or athiazolidinedione. (BYETTA® exenatide injection. Prescribinginformation; DeFronzo, et al. Diabetes Care 28:1092-1100, 2005; Kendall,et al. Diabetes Care 28:1083-1091, 2005; Zinman, et al. Ann Intern Med146:477-485, 2007; Buse, et al. Diabetes Care 27:2628-2635, 2004).Despite modest weight loss and improved glycemic control,gastrointestinal (GI) intolerability and the need for twice-dailyinjections often leads to discontinuation (Fineman, et al. DiabetesMetab Res Rev 20:411-417, 2004).

Albiglutide is a GLP-1 receptor agonist developed by fusion of twoDPP-4-resistant human GLP-1 analogs to human albumin (Matthews, et al. JClin Endocrinol Metab. 2008; DOI: 10.1210/jc.2008-1518). Its extendedhalf-life (˜5 days) may allow weekly or less-frequent dosing. This studywas designed to explore a wide range of doses (4-100 mg) and schedules(weekly-to-monthly) to assess glycemic control and adverse eventprofiles. Exenatide was included as an open-label reference to provideclinical perspective for a GLP-1 receptor agonist.

Research Design and Methods Protocol

This phase 2 trial was a prospective, randomized, double-blind,placebo-controlled, parallel group study conducted between April 2007and May 2008 in 118 sites in the United States (n=106), Mexico (n=9),Chile (n=2), and the Dominican Republic (n=1). Men and women ofnon-childbearing potential from 18-75 years of age were eligible forinclusion if diagnosed with type 2 diabetes ≧3 months before screening.Subjects were drug-naïve (diet & exercise) or treated with diet andexercise plus metformin as monotherapy stable for >3 months beforeprescreening (1 week prior to screening visit). Only subjects treatedwith metformin monotherapy were eligible for the exenatide arm(consistent with labeling). Additional inclusion criteria included: BMI≧20 and ≦40 kg/m² and A1C at screening ≧7% and ≦10%.

Exclusion criteria included: any oral diabetes monotherapy (exceptmetformin)≦3 months prior to screening or insulin <1 month prior toscreening and not used for >7 days; pancreatitis <5 years; significantcardiovascular, cerebrovascular, renal or hepatobiliary disease; fastingserum triglycerides ≧800 mg/dL (9 mmol/L) at screening; andhematological profiles considered to be clinically significant. Subjectstaking lipid-lowering medications must have been maintained at the samedose for 3 months prior to enrollment. Prescription or over-the-counterweight-loss drugs were not permitted.

The study protocol was approved by an Institutional Review Board andconducted in accordance with Good Clinical Practice and the Declarationof Helsinki Written informed consent was obtained from all subjects atprescreening. A Data Safety Monitoring Committee of independent expertsassessed safety data on an ongoing basis.

Randomization

Subjects were randomized into 1 of 10 treatment arms: double-blindplacebo (matched to albiglutide arms); albiglutide weekly (4, 15 or 30mg), biweekly (15, 30 or 50 mg) or monthly (50 or 100 mg); or open-labelexenatide (5 μg twice daily for 4 weeks followed by 12 weeks of 10 μgtwice daily). Albiglutide and placebo were administered in thephysician's office over the course of 16 weeks. Subjects receiving 4 mgalbiglutide were given 1.0 mL (4 mg/mL solution). Subjects receiving 15mg, 30 mg, or 50 mg albiglutide were given 0.32 mL, 0.65 mL, or 1.0 mL(50 mg/ml solution). Subjects receiving 100 mg albiglutide were given2×1.0 mL injections (50 mg/mL solution, >1 inch apart). Placebo volumeswere matched to active treatment. Albiglutide/placebo injections weresubcutaneous to the abdomen using 30G needles. Subjects were observedfor at ≧30 minutes to monitor for injection site reactions. Subjectsreceiving exenatide initiated treatment in the physician's office andsubsequently self-administered according to the package insert. After 16weeks, subjects entered an 11-week washout phase primarily to assesssafety and immunogenicity.

Assessments On-Therapy:

A1C and fasting plasma glucose (FPG) measurements were performed atscreening, baseline, and at weeks 2 (FPG only), 4, 5, 7, 8, 9, 12, 15,and 16. Fasting fructosamine, C-peptide, glucagon, insulin and lipidswere measured at baseline and weeks 8, 12, and 16. β-cell function wascalculated using homeostasis model assessment (HOMA) (Matthews, et al.Diabetologia 28:412-419, 1985).

Adverse event assessments and safety analyses (includingelectrocardiograms, vital sign measurements and physical exams) wereconducted throughout the study. Nausea and vomiting were monitored foroccurrence and duration. Immunogenicity assessments were performed withsamples taken at baseline and at weeks 1, 4, 8, and 12, and 16. Sampleswere screened for anti-albiglutide antibodies via ELISA (Matthews, etal. J Clin Endocrinol Metab. 2008; DOI: 10.1210/jc.2008-1518). Plasmasamples were collected to characterize the pharmacokinetics (PK) ofalbiglutide (quantified by ELISA at baseline and weeks 4, 5, 7, 8, 9,12, 15, and 16). Population PK analysis was performed using a nonlinearmixed-effect modelling approach with NONMEM software (Icon DevelopmentSolutions, Ellicott City, Md.).

11-Week Washout:

Immunogenicity assessments were examined at weeks 20, 23, and 27; A1Cand FPG were obtained at weeks 17, 18, 20, 23, and 27; fastingfructosamine, C-peptide, glucagon, insulin, and lipid profiles wereobtained at weeks 20 and 27; and albiglutide concentrations wereobtained at weeks 17, 18, 20, 23, and 27.

Statistical Analysis

The primary objective was to evaluate the dose response of albiglutidefor safety and efficacy. With 30 subjects planned in each treatment arm,a 2-sided 95% confidence interval for each treatment group mean responsehad a half-width of 0.36% on the A1C measurement scale, assuming astandard deviation (SD) of 1.0%.

The primary efficacy endpoint was change from baseline A1C at week 16 vsplacebo across different doses within each schedule (weekly, biweekly,and monthly). The primary analysis was an ANCOVA model with main effectsfor treatment group and prior metformin therapy, adjusting for baselineA1C. Dose response was evaluated using contrasts within the ANCOVA modelframework. Pairwise comparisons were performed in the same ANCOVA model.Secondary endpoints were analyzed similarly. Responder analysis andincidence of hypoglycemia were summarized by group statistics. No formalstatistical comparisons vs exenatide (open-label) were conducted. Safetyand tolerability data were collected categorically.

Comparisons were made on the intent-to-treat population, defined as allrandomly assigned subjects with at least one post-baseline assessment ofthe primary endpoint, using last-observation carried forward. The safetypopulation included all randomized patients who received at least onedose of any medication after being randomized. An interim analysis wasconducted at 8 weeks for administrative purposes by an independentstatistical analysis group; blinding was retained for studyinvestigators and study personnel with daily operational responsibility.No formal interim inferential hypothesis testing was conducted, thestudy was not terminated early, nor changed based on the result of theinterim analysis.

Patients were also assessed for reduction in fasting plasma glucose(FPG) and HbA1c reduction based on background metfomin (MET) treatment.

Results Subject Disposition and Baseline Characteristics

A total of 774 subjects were screened. Of 361 subjects randomized, 356(mean age 53 years, BMI 32.1 kg/m²) received treatment and were includedin the safety analysis; 345 subjects were included in the efficacyanalysis, and 255 completed the 16-week trial. Withdrawal rates weresimilar across groups, the most frequent reasons for withdrawal wereadverse events. The most frequent adverse events (occurring in >1patient) leading to withdrawal included hyperglycemia (0-11.8%),gastrointestinal events (0-11.4%), events associated with injection site(0-9.7%) and hypertriglyceridemia (5.7%). Other reasons for withdrawalincluded loss to follow-up, protocol violations, and voluntarywithdrawal.

Baseline demographics and characteristics were comparable across groups.Mean duration of diabetes was 5 years, and baseline A1C levels (mean8.0%) were evenly distributed across arms. A similar proportion ofsubjects receiving placebo or albiglutide were drug-naïve (25.7-34.4%)or receiving metformin as monotherapy. All subjects receiving exenatidewere on background metformin monotherapy. The groups were similar interms of race and ethnicity (43.8-64.5% white; 87.1% and 12.9% ofsubjects were from US and Latin American clinics, respectively), fastingglucagon (range 94.4-108.9 ng/L), and rates of dyslipidemia,hypertension, and coronary artery disease (ranges: 50.0-80.0%;47.1-67.6%; and 0-15.2%, respectively).

Efficacy

After 16 weeks, albiglutide significantly reduced A1C in a generallydose-dependent manner within each dose schedule (Table 1, FIG. 2A). MeanA1C reductions from baseline in subjects receiving highest dose in eachtreatment schedule were −0.87%, −0.79% and −0.87% for 30 mg weekly, 50mg biweekly and 100 mg monthly, respectively, versus placebo (−0.17%) orexenatide (−0.54%). The A1C reductions (based on ANCOVA model) for thehighest doses compared with placebo were statistically significant: 30mg weekly −0.62% (95% CI −1.03, −0.22), P=0.003; 50 mg biweekly −0.57%(95% CI −0.96, −0.19), P=0.003; and 100 mg monthly −0.60% (95% CI,−0.99, −0.22) P=0.002. As expected, numerically greater reductions inA1C were observed in subjects with baseline A1C≧8.5%.

At week 16, A1C and FPG were reduced dose-dependently within allalbiglutide dosing schedules. A1C was similarly reduced by albiglutide30 mg weekly, 50 mg biweekly and 100 mg monthly (−0.9, −0.8 and −0.9%;respectively, p<0.005) vs PLACEBO (−0.2%). Results are summarized inTables 1 and 2. HbA1c and American Diabetes Association glycemic targetmean change from baseline are shown for all groups in FIG. 2 and FIG. 3.

TABLE 1 FPG A1C < Dose (n) A1C (%)^(†) (mmol/L)^(†) 7.0% (%) Placebo(51) −0.17 ± 1.0  −0.10 ± 2.9 20 Exenatide 5/10 μg (35) −0.5 ± 0.9 −0.80± 2.5 35.3 ALB 4 mg (35) −0.1 ± 1.2 −0.47 ± 3.1 17.6 weekly 15 mg (35)−0.5 ± 0.7 −0.72 ± 1.7 35.3 30 mg (31)  −0.9 ± 0.7*  −1.44 ± 2.0* 51.7ALB 15 mg (33) −0.6 ± 1.0 −1.28 ± 2.4 26.7 biweekly 30 mg (32)  −0.8 ±1.0*  −1.58 ± 2.1* 50.0 50 mg (35)  −0.8 ± 1.0*  −1.32 ± 3.5* 52.9 ALB50 mg (35) −0.6 ± 1.0 −0.72 ± 2.8 22.9 monthly 100 mg (34)  −0.9 ± 0.9* −1.22 ± 3.5* 48.4 *p < .05 vs PBO; ^(†)Mean ± SD (Δ From BL)

TABLE 2 Change from baseline in glycemic parameters at 16 weeksExenatide^(†) Albiglutide Twice Daily Weekly Placebo 5 μg to 10 μg 4 mg15 mg 30 mg N 51 35 35 35 31 Baseline A1C (%) 7.9 ± 0.9   7.9 ± 0.9 8.0± 1.0   8.1 ± 0.9   8.0 ± 0.9 Mean ± SD Δ A1C at 16 weeks vs −0.17 ±1.01   −0.54 ± 0.91 −0.11 ± 1.16   −0.49 ± 0.74 −0.87* ± 0.65  baseline,% ± SD Baseline FPG, 10.0 ± 3.8    9.4 ± 2.4 10.8 ± 3.8    9.7 ± 2.9  9.5 ± 3.1 mmol/L ± SD Δ FPG at 16 weeks vs −0.10 ± 2.90   −0.80 ± 2.48−0.47 ± 3.12   −0.72 ± 1.68 −1.44* ± 2.03  baseline, % ± SD AlbiglutideBiweekly Monthly 15 mg 30 mg 50 mg 50 mg 100 mg N 33 32 35 35 34Baseline A1C (%) 8.2 ± 1.0   8.0 ± 1.0 8.0 ± 0.7   7.9 ± 0.8   8.0 ± 1.0Mean ± SD Δ A1C at 16 weeks vs −0.56 ± 0.97   −0.79* ± 0.98  −0.79* ±1.04    −0.55 ± 1.01 −0.87* ± 0.87  baseline, % ± SD Baseline FPG, 10.2± 2.7    9.5 ± 3.3 10.1 ± 3.2    9.3 ± 2.7   9.7 ± 3.8 mmol/L ± SD Δ FPGat 16 weeks vs −1.28 ± 2.43   −1.58* ± 2.06  −1.32* ± 3.52    −0.72 ±2.77 −1.22* ± 3.50  baseline, % ± SD FPG, fasting plasma glucose; A1C,glycosylated hemoglobin; SD, standard deviation. *p < 0.05 vs placebo^(†)Exenatide was used to provide clinical reference; no statisticalanalyses were conducted.

Weight loss (−0.9 to −1.8 kg) was observed with albiglutide. Documentedhypoglycemia was not increased with albiglutide; most frequentlyreported AEs included nausea, vomiting and headache. Lowest incidence ofgastrointestinal AEs was in those receiving weekly albiglutide 30 mg. Nosubject experienced pancreatitis. Most skin reactions were small andlocalized to injection site. Positive anti-ALB antibody occurred in 8subjects (2.5%), including 1 on PLACEBO and 2 at baseline.

The proportion of subjects achieving ADA target for glycemic control(A1C<7.0%) at week 16 increased with increasing doses within each doseschedule; similar proportions of subjects achieved A1C targets at thehighest albiglutide dose among the three schedules. Accordingly, moresubjects receiving albiglutide 30 mg weekly (52%), 50 mg biweekly (53%),and 100 mg monthly (48%) achieved A1C<7.0%, compared with 20.0% and35.3% of subjects receiving placebo and exenatide, respectively (FIG.2B).

The time course of albiglutide-induced changes in FPG demonstrated thateach dosing schedule of albiglutide elicited a dose-dependent reductionin FPG over 16 weeks, with no changes in FPG observed in subjectsreceiving placebo. Rapid reductions in FPG were observed, with FPGreduction at the 16-week endpoint similar for each of the highest doses(FIG. 3C). Statistically significant reductions were seen for FPGchanges from baseline compared with placebo at week 16 [−1.38 (P=0.01),−1.16 (P=0.03), and −1.17 (P=0.02) mmol/L for 30 mg albiglutide weekly,50 mg biweekly and 100 mg monthly doses, respectively]. The 4 mg and 15mg weekly dose regimens of albiglutide reduced FPG but were lesseffective. Notably, the greatest fluctuations in FPG over time wereobserved in subjects receiving the monthly dosing regimen (FIG. 3C).Exenatide was associated with a relatively consistent FPG profile overtime that was numerically less than FPG reductions seen with the highestdoses of albiglutide (FIG. 3C).

Neither fasting insulin nor glucagon levels were consistently orsignificantly altered. Small improvements in β-cell function (assessedby HOMA-B) were noted in subjects receiving albiglutide.

There was no significant difference in weight reduction among groups. Aconsistent trend in weight reduction was noted, with average weight lossranging from −1.1 to −1.7 kg in subjects receiving albiglutide at thehighest dose regimens in each timing schedule. These reductions werenumerically greater than those receiving placebo (−0.7 kg), but lessthan weight loss with exenatide (−2.4 kg). Albiglutide and exenatidetended to reduce mean systolic and diastolic blood pressure, but did notsignificantly change the plasma lipoprotein profile.

Across albiglutide groups and placebo, 65.6-74.3% received backgroundmetformin (MET). After 16 weeks, background MET subjects experiencedfasting plasma glucose (FPG) reductions of −1.26, −2.10, −1.80 and −0.07mmol/L for the 30 mg weekly, 50 mg biweekly and 100 mg monthly doses ofalbiglutide and placebo, respectively, vs −1.44, −1.32, −1.22 and −0.10mmol/L, respectively, for the overall population. Exenatide decreasedFPG by −0.80 mmol/L.

In MET subjects, the highest albiglutide doses in each dose schedulesignificantly reduced HbA1c similarly over 16 weeks: albiglutide 30 mgweekly, −0.78%; albiglutide 50 mg biweekly −0.83%; and albiglutide 100mg monthly −0.77% vs placebo (−0.05%, p<0.05); exenatide reduced HbA1cby −0.54%. In the overall population, HbA1c was reduced −0.87, −0.79 and−0.87% by albiglutide 30 mg weekly, 50 mg biweekly and 100 mg monthlydosing, respectively, vs placebo (−0.17%, p<0.005).

Among MET patients, HbA1c<7% was achieved by 43%, 50% and 46% ofsubjects receiving albiglutide 30 mg weekly, 50 mg biweekly and 100 mgmonthly, respectively, vs 15% with placebo and 35% in the exenatidegroup. Weight loss with albiglutide was observed in both the MET group(−0.4 to −2.1 kg) and in the overall population (−0.9 to −1.8 kg).Documented hypoglycaemia was not increased with albiglutide, and mostcommonly reported AEs were gastrointestinal events. Rates of nausea orvomiting in MET subjects were 18.2%, 47.8% and 56.5% of patientsreceiving 30 mg weekly, 50 mg biweekly and 100 mg monthly doses ofalbiglutide, respectively, vs 29.0%, 54.3% and 55.9% in the overallstudy population. Nausea or vomiting was experienced by 45.7% in theexenatide group.

Safety and Tolerability

The percentage of patients reporting at least 1 adverse event wassimilar across groups (67-85%). The most frequently reported adverseevents included nausea (11.8-54.3%), vomiting (0-41.2%), headache(5.9-23.5%), dizziness (5.7-14.3%), nasopharyngitis (5.7-11.4%), backpain (0-14.3%), influenza (0-9.7%), upper respiratory tract infections(0-15.2%), and local skin reactions (2.9-28.6%).

The proportion of subjects who experienced nausea and/or vomiting waslower with administration of ≦30 mg albiglutide compared with theproportion of subjects receiving higher doses (within other doseregimens). In the 30 mg weekly arm, 29.0% of subjects experienced nauseaand/or vomiting, compared with 54.3% of subjects in the 50 mg biweeklygroup and 55.9% of patients in the 100 mg monthly group. The percentageof exenatide patients who experienced nausea and/or vomiting also wasnumerically higher (45.7%) than was seen in the 30 mg weekly albiglutidegroups.

Examination of the time course (FIG. 4) of nausea and/or vomitingrevealed that the proportion of subjects experiencing nausea and/orvomiting each week was low in the mg weekly arm (<10%), and declinedover the course of the study, with no reports of nausea or vomitingafter 8 weeks (FIG. 4C). Although the proportion of nausea and/orvomiting in patients receiving the albiglutide 50 mg biweekly dose wasgreater than in subjects receiving albiglutide 30 mg weekly, theincidence of these adverse events also declined over the study period(FIG. 4D). Subjects receiving the 100 mg monthly dose of albiglutidealso experienced higher rates of nausea and/or vomiting, with peakincidence occurring following each monthly dose administration. Theoverall rate was higher for 100 mg monthly than for any otheralbiglutide group (FIG. 4E). The incidence of nausea and/or vomitingwith exenatide reached 20% by week 2, incidence increased at week 5 to apeak incidence of 29% (due to label-based titration) and also declinedover the study period (FIG. 4B).

Other adverse events were less common than GI-related events and weresimilar across groups, with no dose-dependent trends. Documentedhypoglycemia was not increased with albiglutide (0-3.1%) relative toplacebo (3.9%) and exenatide (2.9%). Cardiac-related adverse events (8subjects) were distributed across groups with no dose-dependent trends.No episodes of pancreatitis were reported.

An 11-week post-treatment washout period was included to monitordevelopment of anti-albiglutide antibodies. A total of eight (2.5%)subjects confirmed positive at least once for anti-albiglutideantibodies in the placebo and albiglutide arms after their baselinemeasurement. However, 2 subjects tested positive prior to albiglutidetreatment (1 each in the 4 mg weekly and 15 mg weekly arms) and 1subject received placebo. The remaining 5 albiglutide-positive subjectswere detected in the albiglutide weekly and biweekly arms. Theappearance of anti-albiglutide antibodies was largely transient, with 1subject remaining positive at week 27. Antibodies were non-neutralizing,low-titer, and in 4 of the 5 subjects showed cross-reactivity withGLP-1. There was no obvious association between presence ofanti-albiglutide antibodies and either efficacy or safety.

Injection site local skin reactions were observed in the study, most ofwhich were small, localized to injection site, and were more common inthe albiglutide groups (2.9-28.2%) compared with placebo (5.9%) andexenatide (2.9%). Injection site reactions tended to occur once/personin subjects receiving 30 mg albiglutide doses, and approximatelytwice/person in subjects receiving higher albiglutide doses. None of theskin reactions was associated with positive IgE antibodies orneutralizing antibodies. The skin reactions did not worsen upon repeateddosing and did not appear to be dose related. No systemic allergicreactions attributable to albiglutide were observed.

Pharmacokinetics

Albiglutide exhibited a half-life of ˜5 days. Steady state levels ofalbiglutide were reached within ˜4-5 weeks of the first dose. Greaterpeak/trough fluctuations in circulating albiglutide concentrations wereobserved with the less-frequent administration of higher albiglutidedoses.

CONCLUSIONS

In this study, the dose- and time-dependent effects of albiglutide, along-acting GLP-1 receptor agonist, were evaluated to identify potentialdose regimens for future studies. Within each dose schedule, albiglutideappeared to be associated with dose-dependent A1C reductions that weresignificantly different from placebo. Maximum doses used for eachschedule (albiglutide 30 mg weekly, 50 mg biweekly, and 100 mg monthly)elicited similar reponses in A1C, providing meaningful reductions withinthe range ˜0.8-0.9% from a mean baseline A1C of 8.0%.

Albiglutide also significantly reduced FPG at week 16 compared withplacebo. FPG reductions were observed at the time of the firstassessment (2 weeks post-dose). In a previous study, FPG reductions wereobserved as early as 2 days following a single dose.

Variability in glycemic response appeared to be related to circulatingconcentrations of albiglutide. With a half-life of ˜5 days and at dosessufficient to achieve consistent therapeutic response (i.e., 30 mg),weekly dosing provided consistent FPG reduction; greater fluctuations inFPG were observed following biweekly or monthly dosing despite similarA1C reductions.

Albiglutide 30 mg weekly dosing elicited steady and consistentimprovement in FPG reductions with a nausea and vomiting profile morefavorable than the reference comparator, exenatide. When dosed biweekly,50 mg albiglutide also improved glycemic indices, but with higher GIadverse event rates possibly related to the higher initial dose. In alldosing schedules, rates of nausea and vomiting declined over time. Anescalating-dose titration for the biweekly regimen might have resultedin a lower frequency of GI events and will be tested in future studies.However, when dosed monthly, albiglutide (50 or 100 mg) did not appearto produce stable FPG reductions between dosing and was associated withhigher GI event rates. The increase in FPG fluctuation and GI events inthe biweekly and monthly regimens were most likely due to fluctuationsin albiglutide concentrations resulting from less-frequent dosing. Takentogether, the efficacy and safety profile in this study suggests thatweekly dosing with at least 30 mg albiglutide provides rapid andsustained glycemic control accomanied by favorable GI tolerability.Future studies may be designed to investigate whether a biweeklyschedule could be an attractive maintenance option for patients whorespond and tolerate the initial weekly regimen.

Mechanistically, reasons for differences in the tolerability profile ofalbiglutide and exenatide are unknown but may be due to differences inpharmacokinetics (T_(max) is ˜3 days vs 2.1 hours for albiglutide andexenatide, respectively) that result in a long half-life of ˜5 days anda steady state achieved after 4-5 doses for 30 mg albiglutide weekly.The slow accumulation of albiglutide may ameliorate the GIintolerability often observed with short-acting GLP-1 mimetics. Inaddition, since albiglutide is relatively impermeant to the centralnervous system (Baggio, et al. Diabetes 53:2492-2500, 2004), it may havea more benign profile with respect to nausea and vomiting than doesexenatide.

Weight loss was similar across albiglutide arms and numerically lessthan the exenatide reference arm. However, larger, longer-term studiesare needed to determine the true effect on weight and cardiometabolicparameters.

Immunogenicity of albiglutide was closely monitored owing to thepossible appearance of neutralizing antibodies or development ofimmediate hypersensitivity reactions. In the present study,anti-albiglutide antibodies were detected in 2.5% (n=8) of subjects.However, the observation that positive titers of anti-albiglutideantibodies were detected in 2 subjects at baseline suggests that theimmunogenicity rate may be overestimated. Exenatide, which has ˜50%homology to human GLP-1, (Drucker, et al. Proc Natl Acad Sci USA84:3434-3438, 1987) is associated with treatment-emergent antibodydevelopment following administration with twice-daily (DeFronzo, et al.Diabetes Care 28:1092-1100, 2005; Kendall, et al. Diabetes Care28:1083-1091, 2005; Zinman, et al. Ann Intern Med 146:477-485, 2007;Buse, et al. Diabetes Care 27:2628-2635, 2004) and weekly (Drucker, etal. Lancet 372:1240-1250, 2008) formulations (>40%). Antibody formationmay attenuate efficacy, especially among patients developing high levelsof anti-exenatide antibodies (Drucker, et al. Lancet 372:1240-1250,2008).

There are limitations of this phase 2 dose- and schedule-finding study.First, the number of patients in each arm is relatively small comparedwith phase 3 studies. Second, relative to the total number of subjects,the drop-out rate was high, due to adverse events, loss to follow-up,and voluntary withdrawals. Third, the duration of active treatment was16 weeks, so full appreciation of the magnitude or durability ofresponse cannot be determined. Finally, no escalating doses were testedfor biweekly and monthly dosing that may have attenuated the frequencyof GI adverse events and fluctuating FPG response.

In summary, albiglutide improves glucose control in a dose-dependentmanner when given weekly and biweekly. Higher monthly doses ofalbiglutide are efficacious, but their use is constrained by the higherfrequency of GI-related adverse events. In conclusion, albiglutideadministered weekly significantly improved glycemic control vs placebo,with an acceptable safety and tolerability profile, modest weight loss,and without increasing the risk of hypoglycemia or immunologicalresponse in subjects with type 2 diabetes. In conclusion, weeklyalbiglutide significantly improved glycemic control with a favorablesafety and tolerability profile; biweekly albiglutide may have potentialas a maintenance option in patients with T2DM.

Albiglutide was effective in patients receiving background MET andprovided numerically greater HbA1c and FPG reductions than didexenatide. Tolerability was most favourable with the 30 mg weeklyalbiglutide group.

Example 2 Gastrointestinal Adverse Event Profile of Albiglutide inSubjects with Type 2 Diabetes

Gastrointestinal (GI) adverse events may limit adherence to GLP-1therapies. The time course of nausea and vomiting (N&V) was assessed ina 16-week randomized, multicenter, double-blind, parallel-group study;356 subjects with type 2 diabetes (T2D) received placebo (PBO),albiglutide (ALB) [weekly (4, 15 or 30 mg), biweekly (15, 30 or 50 mg)or monthly (50 or 100 mg)] or exenatide (Ex, open label) over 16 wks.Combined incidence of N&V in placebo and exenatide was 11.8% and 45.7%,respectively. Combined incidence of N&V was reported by 29%, 54.3% and55.9% of subjects receiving albiglutide 30 mg weekly, 50 mg biweekly and100 mg monthly (mean duration: 2.3, 3.3 and 5.8 d). Incidence of N&V waslower with more-frequent, smaller albiglutide doses vs less-frequenthigher albiglutide doses. All 30 mg weekly events were mild; >90% of 50mg biweekly or 100 mg monthly were mild/moderate. N&V correlated withalbiglutide exposure, and decreased over time. N&V are summarized inTable 3 below.

TABLE 3 ALB 30 mg ALB 50 mg ALB 100 mg Week PBO Exenatide weeklybiweekly monthly 1 2/0  14.3/2.9 6.5/3.2 25.7/5.7 29.4/17.6 2 3.9/0     20/5.7 6.5/0   12.1/0   15.2/3    3 0/0 15.2/3  9.7/3.2 18.2/6.16.3/3.1 4 2/0  12.5/3.1 6.7/0    3/0 3.1/3.1 5 2/0    29/9.7 3.3/3.321.9/9.4 41.9/22.6 6 0/0    29/6.5 6.9/0    6.3/3.1 6.5/3.2 7 0/0 25.8/6.5 6.9/3.4 12.5/6.3 3.2/3.2 8 0/0 19.4/0  3.4/3.4 12.9/3.23.2/3.2 9 2.1/2.1 16.1/0  0/0 16.1/6.5 25.8/19.4 10 0/0  19.4/3.2 0/010.3/0   6.9/3.4 11 0/0 6.5/0 0/0 7.7/0  7.1/3.6 12 0/0 6.5/0 0/0  4/43.7/3.7 13 0/0 6.5/0 0/0  8/8 18.5/11.1 14 2.3/0   3.2/0 0/0  4/0 0/0 152.4/0   3.2/0 0/0  0/0 0/0 16 0/0   0/0 0/0  0/0 0/0 Data = N&V/vomitingalone, %

FIG. 4 shows a “Time Course of Nausea and Vomiting as Adverse Events.”The percentage of subjects experiencing vomiting with or without nausea(gray bars) or nausea (white bars) adverse events during each week ofthe 16 Week trial is shown for A) placebo, B) exenatide, C) albiglutide30 mg weekly, D) albiglutide 50 mg biweekly and E) albiglutide 100 mgmonthly.

In conclusion, weekly 30 mg albiglutide shows a favorable N&V profile vsother albiglutide regimens and Exenatide.

Example 3 Safety, Pharmacokinetics and Pharmacodynamics of Albiglutidein Japanese Subjects with Type 2 Diabetes: A Phase I/II Study

Albiglutide (ALB) is a long-acting GLP-1 mimetic shown to improveindices of glycemia in Caucasian/Hispanic populations when dosed weekly,once every other week (biweekly) and monthly. Pharmacokinetics,pharmacodynamics and safety/tolerability of albiglutide in Japanesesubjects with type 2 diabetes (T2D) were assessed in this 28 day,single-blind, randomized, placebo (PBO)-controlled study; 40 subjects(mean 54.5 y, BMI 24.5 kg/m², A1C range 6.3-10.3%) were given (sc,abdomen) albiglutide: 15 mg or 30 mg once-weekly; 50 mg biweekly; or 100mg monthly. Albiglutide was generally well-tolerated; GI events werecomparable to placebo in all doses except 100 mg monthly and werenumerically lowest in the 30 mg weekly group. In the 100 mg monthlygroup, most common AEs were flatulence (n=3, 38%), vomiting (n=3, 38%)and nausea (n=2, 25%). No serious AEs were reported. Albiglutide had aplasma t½ of 5.3 d, CL/F of 68.7 mL/hr, and V/F of 12.6 L. FPG andweighted mean AUC0-4 glucose were improved as early as day 3. At week29, all doses of albiglutide except 100 mg monthly dose showed asignificant change from baseline compared with PLACEBO for FPG andAUC0-4 glucose. Albiglutide also significantly reduced A1C at all doseson day 29 and 43. Results are summarized in Table 4 below.

TABLE 4 Albiglutide Dose PBO-adjusted change 15 mg weekly 30 mg weekly50 mg biweekly 100 mg monthly from baseline (LS Means (n = 8) (n = 8) (n= 8) (n = 8) Difference) FPG, mg/dL −34.5* −35.6* −31.3* −13.2 (day 29)AUC⁰⁻⁴ glucose, mg/mL −51.6* −64.5* −45.2* −25.9 (day 29) A1C, % (day29/43) −0.58*/−0.87* −0.57*/−0.78* −0.63*/−0.79* −0.51*/−0.59* *P < .05vs PBO

In conclusion, weekly/biweekly albiglutide significantly improvedglycemic control with a favorable safety and tolerability profile inJapanese subjects with T2D.

Example 4 Albiglutide Administered in Combination with Insulin Glargine

Albiglutide is administered as a weekly subcutaneously injected dose ofalbiglutide in combination with insulin glargine as compared with thecombination of insulin glargine and preprandial lispro insulin insubjects with type 2 diabetes. Subjects with a historical diagnosis oftype 2 diabetes who are inadequately controlled despite the use ofinsulin glargine or other intermediate- or long-acting insulins for >/=6months but <5 years, with or without oral antidiabetic medications, whoare unable to achieve a glycosylated hemoglobin value of <7% will berecruited into the study. Subjects must also be willing and capable ofpursuing an intensive regimen of both basal and preprandial insulin.

1. A method of treating diabetes in a human comprising administering to said human a an initial dose of a pharmaceutical composition comprising about 30 mg of a polypeptide comprising the amino acid sequence set forth in SEQ ID NO:1.
 2. The method of claim 1, wherein said pharmaceutical composition is administered subcutaneously.
 3. The method of claim 1, wherein said pharmaceutical composition is administered as a subcutaneous injection selected from the group of at least one 0.32 mL injection, at least one 0.65 mL injection, and at least one 1.0 mL injection.
 4. The method of claim 1, wherein said pharmaceutical composition reduces serum glucose in said human.
 5. The method of claim 1, wherein said human has type II diabetes mellitus.
 6. The method of claim 1 wherein administration of said pharmaceutical composition causes weight loss in said human.
 7. The method of claim 1, wherein said pharmaceutical composition is administered subcutaneously in the leg, arm, or abdomen of said human.
 8. The method of claim 1 wherein said human has received metformin previous to receiving said pharmaceutical composition.
 9. The method of claim 1 wherein said pharmaceutical composition is administered as monotherapy.
 10. The method of claim 1 wherein said pharmaceutical composition is co-administered with at least one second hypoglycaemic agent.
 11. The method of claim 10 wherein said at least one second hypoglycaemic agent is selected from: a second GLP-1 agonist, exendin, sulfonylurea, meglitinide, acetohexamide, chlorpropamide, tolazamide, glipizide, gliclazide, glibenclamide (glyburide), gliquidone, glimepiride, DPP-IV inhibitor, glitazone, thiazolidinedione, rosiglitazone, pioglitazone, pPAR agonist, metformin, α-glucosidase inhibitor, insulin glargine and insulin.
 12. The method of claim 1 wherein said pharmaceutical composition is administered to said human at an initial dose of 30 mg/week of a polypeptide comprising the amino acid sequence set forth in SEQ ID NO:1 and subsequently titrated up to a dose comprising 50 mg/week of a polypeptide comprising the amino acid sequence set forth in SEQ ID NO:1. 